Anti-Tim3 (T Cell Immunoglobulin and Mucin Domain-containing 3, T Cell Immunoglobulin Mucin 3, TIM3,

T cell immunoglobulin mucin-3 (Tim3) is a member of the T cell Immunoglobulin- and Mucin-domain-containing family of type I membrane glycoproteins that regulate autoimmune and allergic disease. Tim3 is selectively expressed on Th1 cells and interacts with galectin-9. It negatively regulates Th1 responses and affects macrophage activation. The 280aa mature human Tim3 contains a V-type Ig-like domain that shows multiple polymorphisms, followed by a mucin-like domain in the 171aa extracellular region, which shares 60% aa identity with the mouse Tim3 ECD. One splice variant is truncated within the mucin domain and presumably is secreted. Applications: Suitable for use in Flow Cytometry. Other applications not tested. Recommended Dilution: Flow Cytometry: 10ul labels up to 1x10e6 cells. Validated using whole blood monocytes., Optimal dilutions to be determined by the researcher. Flow Cytometry Protocol: 1. Cells may be Fc-blocked with 1ug of human IgG/10e5 cells for 15 minutes at RT. Do not wash excess blocking IgG from this reaction., 2. After blocking, add 10ul of T5469-92F to up to 1x10e6 cells and incubate for 30 minutes at RT, 3. Remove unbound antibody by washing the cells twice in Flow Cytometry Staining Buffer (Catalog #F4568-10). Note that whole blood requires a RBC lysis step at this point using Flow Cytometry Human Lyse Buffer., 4. Resuspend the cells in Flow Cytometry Staining Buffer for final flow cytometric analysis. As a control for this analysis, cells in a separate tube should be treated with APC-labeled rat IgG2A antibody. Storage and Stability: Store product at 4°C in the dark. DO NOT FREEZE! Stable at 4°C for 12 months after receipt as an undiluted liquid. Caution: APC conjugates are sensitive to light. For maximum recovery of product, centrifuge the original vial prior to removing the cap.